DK3022300T3 - : Pullulanasekimærer og polynukleotider, som koder for dem - Google Patents

: Pullulanasekimærer og polynukleotider, som koder for dem Download PDF

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DK3022300T3
DK3022300T3 DK14738827.6T DK14738827T DK3022300T3 DK 3022300 T3 DK3022300 T3 DK 3022300T3 DK 14738827 T DK14738827 T DK 14738827T DK 3022300 T3 DK3022300 T3 DK 3022300T3
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DK14738827.6T
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Tomoko Matsui
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Novozymes As
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2451—Glucanases acting on alpha-1,6-glucosidic bonds
    • C12N9/2457—Pullulanase (3.2.1.41)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
    • C12C7/00—Preparation of wort
    • C12C7/04—Preparation or treatment of the mash
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/02—Monosaccharides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/12—Disaccharides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01041—Pullulanase (3.2.1.41)
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/10—Biofuels, e.g. bio-diesel

Claims (10)

1. Poiypeptid med puflulanaseaktivitet, som er valgt fra gruppen bestående al: ίΛ..<Ιβ.Γ udviser mindst 90% sekvensidentitet med aminosyrerne 34 ti! 881 pige SEP ID 140: 9, eller et poiypeptid, der udviser mindst 95% sekvensidentitet med aminosyrerne 34 ti! 881 ifølge SEO ID NO: 11; og hvor den relative aktivitet ved 67X sammenlignet med ved S5X er højere i sammenligning med den relative aktivitet al enzymerne, der er beskrevet som SEO ID NO: 1 og SEQ IDNG: 3, som målt ved anvendelse af PAH8AH~ assiyef ved pH 5,0,
2. Poiypeptid ifølge krav 1, søm udviser mindst 91%, mindst92%, mindst 93%, mindst 94%, mindst 95%, mindst 96%, mindst 97%f mindst 98%,mindst 99% e!!er100%sekvensidentiiei med aminosyrerne 34 til 861 ifølge SEQ ID NO: 9 3 Poiypeptid iføige krav 1, somiudviser mindst 96%. mindst 97%, mindst 96%, mindst 99% eller 100% sekvensidentitet med aminosyrerne 34 til 861 ifølge SEQ ID NO: 11.
4. Sammensætning, søm omfatter poiypepiidet ifølge ethvifkeisom helst af kravene 1-3,
5. Sammensætning ifølge krav 4, som omfatter et eller flere enzymer valgt fra gruppen bestående af: glucoamyiase, alta-ømylase, betaæmylase øg protease.
8. Sammensætning ifølgeetlwiikei som helst af kravene 4 og 5, som omfatter følgende enzymer enpuliuianas©;,engiue©amyiasé,en sliæarnylåse og enprotéasé; eller enprilulønase, en aifæarnylase og en profease;eiSeren puliulanase, englucoamyiaseog en alfa-amylase; eller en puiluianase og en beta-amylase.
7. Sammensætning ildfge et hvilkef som héist al kravene 445, hvor sammensætningen omfatter en puiluianase og eri glucoamyiase og eventueit en alfaæmyiase, og hvor puSlulanasen er valgt blandt et poiypeptid.som udviser mindst 93%: mindst 94%, mindst 95%, mindst 98%, mindst 97%^ mindst 98%, mindsi 99% sokvensidentitet med aminosyrerne 34 til 861 ifølge SEQ iD NO; 11, og glucoamyfasen er valgt blandt i) en G/oeophy/torø frabet/m-glucoamylasevariant, som omfatter substitutionerne S95P+A121P i sammenligning med Gioeophylium trabeum-viidfype-glucoamylase-aminosyresekvensen ifølge SEQ ID NO: 18; eller ii) en variant, som udviser mindst 85%, mindst 90%, mindst 91%, mindst 92%, mindst 93%, mindst 94%, mindst 95%, niindst 96%, mindst 98%, min^ SD NO: 18, og aifa-amylasen er valgt blandt: i) en firt/zomucor pus/Z/us-aifa-amyiasevariant med Aspergillus niger~g\ucoamylaseiinker og -stiyélsesbindende domæne f SS!!},som omfatter subststu ttonerne G128D+D143N i sammenligning med den hybride Rhizommør pua/fcs-alfø-amyiaseaminosyresekvens iføige SEQ ID NO: 14; eller it) en variant, som udviser mindst 85%, mindst 90%t mindst91%, mindst 92%, mindst 93%, mindst 94%, mindst 95%, mindst 96%, mindst 97%, mindst 98%, mindst 99% sekvensidentitet med polypeptidet ifølge SEQ IDJNO; 14; S Ariverideise af et poiypéptid iføige et hvilket som helst af kravene 1-3 til fremstilling af en sirup og/elier et fermenteringsprodukt ud fra et stivelsesholdigt mafertafø, Fremgangsmåde til fremstiiiing af et fermenteringsprodukt lid fra stivelsesholdigt: materiale, hvilken fremgangsmåde omfatter følgende trin; (a) at gøre stivelsesholdigt matenaSe flydende i nærværelse af en aifa-amyiase; :;$$) at forsukre materialet, som er gjort flydende, i nærværelse afenglucoamylase; og i fe) at fermentere med en fermenterende organiémé; hvor trin (a) og/eller trin (b) udføres i nærværelse af et i^lket som helst af kravene 1 -3. 10 Fremgangsmåde til fremstilling af et fermenteringsprodukt ud fra stiyelsiiShojdigt materiale hvilken fremgangsmåde omfatter følgende trin; (a) at forsukre stivelsesholdigt materiale ved en temperatur, som er lavere end deri laveste gelatineringstemperatur for det stivelsesholdige materiale; og (b) at fermentere med en fermenterende organisme, hvor triri få) udføres ved anvendelse af mindst en giucoamylase og et polypeptid iføige et hvilket som helst af kravene :1--3.
11, Fremgangsmåde til fremstilling af et sirupprodukt ud fa stivelsesholdigt materiale; hvilken ffømgangsrøåie omfatter følgende trin: fa) at gøre stivelsesholdigt materiale flydende ! nærværelse af en alfa-amyiase; (b) at forsukre materialet, som er gjort flydende.inasrværelse af en glueosmylase, hvor pulSuianasen iføige et hvilket som helst af kravene 1-3 er til stede under irintfe),
12, Fremgangsmåde ifølge et hvilket som helst af kravene 9-11, hvor det stivelsesholdige materiale er valgt blandt byg. bønnen cassava, kom,:;majs,.; mlio, ærter, kartofler, ris, rug, sago, sorghum, søde kartofler, tapioka, hvede og hele kerner eller en hvilken som helst blanding deraf.
13, Polynukieotid, som koder for polypeptide! ifølge et hvilket som helst af kravene 1-3.
14, NukSeinsyrekonstruktion ellerekspressionsvektor, som omfatter poiynukieotidet ifølge |y^v 13, hvilket poiynukleotid er operativt forbundet tii en eller flere kontrolsekvenser, der styrer fremstillingen af polypeptidet i en ekspressionsvært.
15, Rekombinant værtscelle, som omfatter polynukleobtiet ifølge krav 13, hvilket poiynukleotid er operativt forbundet til en eller tiere kontrolsekvenser, der styrer fremstil i ingen af polypeptidet.
DK14738827.6T 2013-07-17 2014-07-11 : Pullulanasekimærer og polynukleotider, som koder for dem DK3022300T3 (da)

Applications Claiming Priority (2)

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EP13176791 2013-07-17
PCT/EP2014/064914 WO2015007639A1 (en) 2013-07-17 2014-07-11 Pullulanase chimeras and polynucleotides encoding same

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US (1) US9951323B2 (da)
EP (1) EP3022300B1 (da)
CN (1) CN105339494B (da)
AR (1) AR096920A1 (da)
CA (1) CA2915336C (da)
DK (1) DK3022300T3 (da)
HU (1) HUE040545T2 (da)
WO (1) WO2015007639A1 (da)

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MX338068B (es) 2010-04-14 2016-04-01 Novozymes As Polipeptidos que tienen actividad de glucoamilasa y polinucleotidos que codifican los mismos.
ES2673940T3 (es) 2010-12-22 2018-06-26 Novozymes North America, Inc. Proceso para producir productos de fermentación a partir de materiales que contienen almidón
EA201490216A1 (ru) 2011-07-06 2014-07-30 Новозимс А/С Варианты альфа-амилазы и кодирующие их полинуклеотиды
IN2014CN02469A (da) 2011-09-06 2015-06-19 Novozymes As
MX351762B (es) 2011-10-11 2017-10-26 Novozymes As Variantes de glucoamilasas y polinucleotidos que las codifican.
IN2014CN03467A (da) 2011-10-11 2015-10-16 Novozymes North America Inc

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CN105339494A (zh) 2016-02-17
US9951323B2 (en) 2018-04-24
CA2915336A1 (en) 2015-01-22
CA2915336C (en) 2024-03-12
AR096920A1 (es) 2016-02-03
EP3022300B1 (en) 2018-07-11
WO2015007639A1 (en) 2015-01-22
EP3022300A1 (en) 2016-05-25
HUE040545T2 (hu) 2019-03-28
US20170051265A1 (en) 2017-02-23
CN105339494B (zh) 2020-04-17

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